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Novelty:We report a one step assay for the parallel quantification of miRNA in tens of thousands of single cells.Our protocol leverages DSN (duplex-specific nuclease) induced signal amplification (Fig.1) [1] and uses pico-liter sized micro-wells, which confine single cells and the enzymatic reaction.We demonstrated our assay with triplexed miRNA detection in K562 cells and a detection limit of miRNA at 5.7 copies per well.